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Research Team
Research Team
Zack Hesselmann
Zack Hesselmann
Major: Biology
Minor: Neuroscience
Jacob Weaver
Jacob Weaver
Doctoral Student
Biology
Abstract/Introduction
Abstract/Introduction
FOXE3 is a transcription factor that plays a crucial role in the early development of the ocular lens. Mutations in FOXE3 across many species (Mouse, frog, human, zebrafish) result in devastating phenotypes such as primary aphakia (loss of the lens), Peters anomaly (attachment of lens to the cornea) and cataracts. However, how FOXE3 mechanically works and how these severe phenotypes arise from FOXE3 mutations remains a mystery. The proposed project will help elucidate how FOXE3 works in early development. I will conduct a temporal assessment of phenotypes resulting from the loss of FOXE3 by comparing eye development in wild-type and FOXE3 knock out (KO) mice at 3 different stages of embryonic development: embryonic day 12, 15 and postnatal day 0 (E12, E15 and P0). We will collect, fix, and embed tissue samples in paraffin. I will section these tissues for histological analyses including: hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) to identify relevant proteins that previous RNA-sequencing data suggest may be altered in the absence of FOXE3. These analyses will play a critical role in identifying target proteins that are under the regulation of FOXE3. By understanding how FOXE3 affects lens structure we will also better understand what exactly FOXE3 does during embryonic eye development. The protein data generated by this project should complement the existing transcriptomic data in a manuscript that we will submit next year. Lastly, this project may inspire other biomedically oriented undergraduate students to do research on topics directly relevant to human development and birth defects.
Research Aims
Research Aims
Aim 1: How does the deletion of FOXE3 affect the structure of the lens at various time points of embryonic development?
Aim 2: Do the changes in RNA transcript expression in FOXE3 knockouts translate to differences at the protein level?
Methods
Methods
Collect mouse tissues samples of FOXE3 knockout (KO) and wildtype (WT) mice at stages E12, E15, and P0
Fix tissue samples in formalin and embed into paraffin
Section paraffin blocks using microtome and put on slides.
Deparaffinization section with Xylene and rehydrate tissue with a series of alcohol washes
For Hematoxylin and Eosin (H&E) staining, we stain for nuclei and cytoplasm to see cell morphology
For immunohistochemistry (IHC), we probe for specific proteins
The following is an image of poster presented at the 2026 Undergraduate Research Forum
Acknowledgements
Acknowledgements
R21 EY031092 Grant- National Eye Institute of Health
G20230315-5759 Grant – Sigma Xi
Office Of Undergraduate Research at Miami University URA
References
References
[1] Robinson M L Lens Fiber Cell Differentiation. In: Darlene A. Dartt, editor. Encyclopedia of the Eye, Vol 2. Oxford: Academic Press; 2010. pp. 543-550.
NACE Career Readiness Competencies
NACE Career Readiness Competencies
Teamwork: Over the course of this project, I have been extremely lucky to have mentors that truly care about my learning and project progress. I didn't realize how important teamwork was in the realm of research until I started actually doing research in the Robinson Lab. There are so many instances during this project that I learned how valuable it is to have good teamworking skills. Coming to a common goal requires effective communication, accountability, and positive working relationships.
Technology: I learned many new laboratory techniques that, before, I had only seen in textbooks. Jacob, my primary mentor, taught me how to perform many of these techniques, but more importantly he took the time to explain the why behind these techniques. For example, if you put most people in a lab with everything they need and a protocol, they will most likely be able to do the experiment. However, without an outstanding mentor, you won't know why you did the steps that you did. Jacob taught me the ins and outs of technological techniques like sectioning, immunohistochemistry, hematoxylin and eosin staining, and imaging.
Critical Thinking: I learned how to apply critical thinking skills during this project. From project planning, to navigating complex protocols, I learned how to make decisions and solve problems. Specially, I remember doing an immunohistochemistry protocol by myself for the first time; there were many instances where I wanted to just call for help, but instead I used my critical thinking skills to figure out quick solutions.
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